tbuckley/cabot-search huggingface dataset:
PB1783: IDENTIFICATION OF RAMAN SPECTROSCOPIC SIGNATURE OF IDH MUTATIONS IN AML IN VITRO MODEL
Authors: P. Laskowska, A. Nowakowska, S. Orzechowska, A. Borek-Dorosz, J. Stolarska, A. Adamczyk, P. Leszczenko, M. Zasowska, A. Szlachetka, M. Szydłowski, E. Białopiotrowicz, P. Juszczyński, M. Barańska, K. Majzner, P. Mrówka
Journal: HemaSphere (2022)

Background: Current acute myeloid leukaemia (AML) laboratory diagnostics is based on genotyping and immunophenotyping. Although these methods revolutionized the way we understand, diagnose and treat myeloid malignancies, it does not free from disadvantages such as cost or lack of functional information on cells state. Raman spectroscopy, the label-free method based on the specificity of light inelastic scattering, has potential to overcome those limitations and enrich diagnostic process with new data. Raman spectra serve as ‘molecular fingerprints’ characterizing biochemical composition of complex cellular structures. We utilized Raman microscopy to analyse the influence of single mutation on Raman spectral images of leukemic cells. Somatic mutations in isocitrate dehydrogenase 1 and 2 (IDH1/2) are recurrently identified, actionable driver events in adult patients with AML. The mutant form of IDH enzymes convert alpha-ketoglutarate to 2-hydroxyglutarate. Accumulation of the latter impairs histone demethylation and leads to the interruption of gene expression. These properties make IDH mutants an ideal target for attempting the label-free identification using Raman spectroscopy. Aims: The aim of the study was to identify Raman spectroscopic signature of IDH oncogene-driven molecular changes in in vitro AML models. Methods: IDH1/R132H, IDH2/R140Q, and IDH2/R172K constructs were retrovirally transduced into HEL and HL-60 cell lines. Cells were imaged using the confocal Raman system WITec Alpha 300. Wild-type forms of genes and empty pMIG plasmid served as control samples. Collected Raman maps were processed with K-means cluster analysis (KMCA) followed by Principal Component Analysis (PCA) and Partial Least Squares–Discrimination Analysis (PLS-DA) performed on averaged spectra of cells and subcellular components. Results: PCA analysis revealed that IDH1/2 mutations (R132H, R140Q and R172K) carrying cells show subtle but significant differences in protein and nucleic acids composition in comparison to control cells. The most significant influence on Raman spectra was observed for IDH1 R132H mutation. Raman features in PCA loadings were also related to proteins and nucleic acids. Using the PLS-DA method, we built a model for distinguishing transgenic IDH1/2-mutated models from IDH wild-type cells. Summary/Conclusion: The results confirm the utility of Raman spectroscopy/microscopy to identify the changes between mutated and wild type forms of IDH1/2 suggesting the potential of optical methods for studying point-mutations in cancer cells and subsequently screening of leukaemia. Further studies to improve and optimise the method and collect more data for machine learning are warranted.
Dressed to impress—and raise awareness
Authors: Alison Shepherd
Journal: BMJ (2022)

Riluzole is the only approved medication that extends survival for patients with amyotrophic lateral sclerosis (ALS). While other potential neuroprotective agents have been evaluated in randomized clinical trials, none has shown unequivocal success and none has been approved by regulatory agencies. Few symptomatic therapies have been tested in ALS. Effectiveness for drugs with modest benefit can be established only through large phase III randomized clinical trials. With numerous potential agents but limited resources, priority should be given to agents that show promise in phase II trials before proceeding to evaluation in phase III trials. In this article, we review drug development in early phase ALS trials and introduce novel designs. First, to maximize the therapeutic potential of the test medication, we need to identify the highest dose that produces a tolerable level of side effects. Second, candidate treatments should be ranked by conducting randomized selection trials between competing new treatments. The selection paradigm adopts a statistical viewpoint different from the hypothesis testing framework in conventional trials. We exemplify this approach by describing a group-sequential selection design developed for a phase II, randomized, multicenter trial of two combination treatments in patients with ALS, and illustrate the sample size reduction from a conventional trial.
PB2230: EFFICACY AND SAFETY OF SINOPHARM VACCINE FOR SARS-COV-2 AND BREAKTHROUGH INFECTIONS IN IRANIAN PATIENTS WITH HEMOGLOBINOPATHIES: A PRELIMINARY REPORT
Authors: M. Karimi, T. Zarei, S. Haghpanah, A. Azarkeivan, M. Naderi, S. Matin, A. Bazrafshan, Z. Zahedi, A. Shirkavand, P. Pishdad, V. De Sanctis
Journal: HemaSphere (2022)

Background: The ongoing COVID-19 pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has led to high morbidity and mortality worldwide. Vaccination against SARS-CoV-2 is a leading strategy to change the course of the COVID-19 pandemic. Aims: Our aim was to investigate the efficacy and side effects of Sinopharm vaccine in patients with hemoglobinopathies in Iran, and the frequency of breakthrough infection after a full course of vaccination. Methods: A multicenter cross-sectional study of 434 patients with hemoglobinopathies (303 β-thalassemia major, 118 β-thalassemia intermedia, and 13 sickle-thalassemia) were conducted from March to July 2021 in IRAN. All patients have received the first dose of China Sinopharm vaccine and received the second dose of the vaccine 28 days apart. Detection of immunity after vaccination was evaluated by commercial enzyme- linked immunosorbent assay (Pishtazteb ELISA commercial kit), including a surrogate virus neutralization test (sVNT), for detection of SARS-CoV-2 immunoglobulins (IgA, IgM, IgG), total neutralizing antibody (NAb). Results: The mean age of patients was 35.0± 8.5 (from 18 to 70) years and 55.6% were positive for antibody. Overall, 48.2% of the studied population had at least one side effect after vaccination. The most frequent side effects were fever and chills, dizziness, and body pain. A total of 90 (20.7%) vaccinated patients developed breakthrough infections after full Sinopharm vaccination. Disease severity was recorded, and it was classified as mild in 77.8%, moderate in 13.6%, and severe in 7.4% of patients. One 28-year-old woman with β- thalassemia major died eight days after the diagnosis of breakthrough SARS-CoV-2 infection. Summary/Conclusion: No safety concerns were identified in patients who received two doses of Sinopharm vaccine. Its efficacy was not optimal which might be due to lack of its effect on new variations of virus. However, our data show that it can prevent from severity of COVID-19 infection in patients with hemoglobinopathies. The frequency of breakthrough infections after full Sinopharm vaccination support the evolving dynamic of SARS-CoV-2 variants requiring special challenge, since such infection may represent a risk for vulnerable patients.
P1513: LOW VITAMIN D LEVELS ARE ASSOCIATED WITH INCREASED CARDIAC IRON UPTAKE IN THALASSEMIA MAJOR.
Authors: A. Meloni, L. Pistoia, C. Vassalle, S. Maffei, I. Fotzi, R. Rosso, M. Serra, A. Quota, P. P. Bitti, V. Carrai, V. Positano, F. Cademartiri
Journal: HemaSphere (2022)

Background: Decreased vitamin D (VitD) levels stimulate the expression of the transmembrane L-type voltage-dependent calcium channels (LVDCC), which, in addition to calcium, absorb iron. However, there is little and controversial data on the correlation of VitD levels with cardiac iron. Aims: We investigated the association between vitamin D levels and cardiac iron and function in thalassemia major (TM) patients. Methods: We considered 278 TM patients (39.04±8.58 years, 56.8% females), consecutively enrolled in the Extension-Myocardial Iron Overload in Thalassemia Network. Myocardial iron overload (MIO) was quantified using a T2* segmental approach. Left ventricular (LV) function parameters were quantified by cine images. Measurement of serum 25(OH)D was carried out using a chemiluminescent immunoassay. Results: Vitamin D supplements were taken by 61.4% of our patients. VitD levels were deficient (<20 ng/dl) in 107 (38.5%) patients, insufficient (20–30 ng/dl) in 96 (34.5%) patients and adequate (>30 ng/dl) in 75 (27.0%) patients. Patients with deficient vitD levels were significantly younger than patients with adequate (36.96±7.64 yrs vs 41.26±8.19 yrs; P<0.0001) as well as insufficient vitD levels (36.96±7.64 yrs vs 39.63±9.39 yrs; P=0.042) while no difference was detected in male to female ratio. MIO (global heart T2*<20 ms) was significantly more frequent in patients with deficient VitD than in patients with adequate and insufficient VitD levels (Figure). Patients with deficient vitD levels had a significant higher risk of MIO than patients with adequate vitD levels (odds ratio-OR=20.62, 95%CI=2.67-153.72; P=0.004) and patients with insufficient VitD levels (OR=8.49, 95%CI=2.46-29.29; P=0.001). At receiver operating characteristic curve analysis, a vitD level≤ 17.3 ng/dl predicted MIO with a sensitivity of 81.5% and a specificity of 75.3% (P<0.0001). The area under the curve was 0.79 (95% confidence interval: 0.74–0.84). Compared to patients with normal VitD levels, patients with deficient vitamin levels showed significantly higher LV end-diastolic volume index (87.61±19.19 ml/m2 vs 79.17±14.27 ml/m2; P=0.003) and LV mass index (63.39±13.55 g/m2 vs 57.45±12.90 g/m2; P=0.006). The LV ejection fraction tended to be lower in patients with VitD deficiency, but no statistical difference versus the other groups was detected. Image:Summary/Conclusion: In thalassemia major vitamin D deficiency is associated with and increased risk of cardiac iron overload. Periodic and regular assessment of VitD levels can be useful in the prevention not only of bone disorders but also of cardiac iron accumulation and subsequent overt dysfunction.
PB1910: CHRONIC MYELOID LEUKEMIA AND RELATIONSHIP TO SECONDARY SOLID NEOPLASMS
Authors: M. J. Ramos, R. Peixeiro, L. Regadas, C. Pedrosa, P. Seabra, A. Mota, R. Cabral, R. Coutinho, L. Xavier, C. Casais, J. Rodrigues, I. Freitas, C. Gonçalves
Journal: HemaSphere (2022)

Background: Chronic Myeloid Leukemia (CML) is a myeloproliferative neoplasm whose first-line treatment involves Tyrosine Kinase Inhibitors (TKI), that have improved the prognosis of the disease after its introduction, with a gain in the overall survival of these patients. In patients in the chronic phase of the disease, the most common cause of death is associated with other neoplasms, which raises the hypothesis that the observed mortality could be the result of a carcinogenic effect of tyrosine kinase inhibitors. Since studies to date have had contradictory results, this work aims to assess the risk of developing secondary solid neoplasms (NSS) in patients with CML. Aims: This study aims to assess the risk of developing SSN in patients with CML. Methods: Patients with positive BCR-ABL transcripts were analysed in a University Hospital Center between 01/01/2008 and 15/02/2022. Results: Sample of 69 patients (53.6% women) all in the chronic phase at diagnosis. Median age 53 years (23-85 years). In the first therapeutic line, most of them underwent TKI (91.3% underwent Imatinib and 5.7% underwent nilotinib). In the total sample, 69.5% needed only 1 therapeutic line. Eight (11.6%) died, only in 2 cases the death was related to CML. Nine patients (11.9%) developed SSN, a majority men (n=8, 88.9%) with a median age of 69 years (57-74 years) at diagnosis. All of them were diagnosed between 14 and 121 months. The majority of patients had gastrointestinal cancer (n=4, 44.4%), followed by prostate cancer (n=3, 33.3%), head and neck cancer (n=1, 11.1%) and lung cancer (n=1, 11.1%). All were exposed to TKI and Imatinib was common to all. Two patients died due to SSN, corresponding to 25% causes of death in the total study sample. When statistically comparing the occurrence of SSN with the time between the diagnosis of CML and the second neoplasm or the duration of the different therapeutic lines, there was no correlation with statistical significance (p>0.05). Summary/Conclusion: SSN appeared in about 12% of patients with CML. Contrary to what is described in the literature, the sample of patients with SSN is mostly made up of older men. As the risk of developing neoplasms in the general population is greater in the elderly, the development of cancer in these patients can be considered to be similar to that in the general population. The greatest risk for the development of SSN is described in the 1st year after the diagnosis of CML, which was not verified in our study. As limitations of the study the authors highlight the reduced sample as well as the need for a longer follow-up period.
P1660: PERIPROCEDURAL MANAGEMENT IN COMBINED DEFICIENCY OF FACTOR V AND FACTOR VIII: EXPERIENCE FROM A SINGLE CENTER IN SPAIN
Authors: E. FERNANDEZ MELLID, P. MELERO VALENTIN, P. CADAHIA FERNANDEZ, A. DE ANDRES Y JACOB, L. BAO PEREZ, A. ABUIN BLANCO, R. FERREIRO FERRO, A. PELETEIRO RAINDO, M. RODRIGUEZ RUIZ
Journal: HemaSphere (2022)

Background: Inherited combined factor V (FV) and factor VIII (FVIII) deficiency is a rare disorder with typically mild bleeding symptoms. There is limited data about replacement therapy which is usually only required before surgical procedures or traumatic hemorrhages. Aims: Revision of hemostatic treatment and bleeding control in FV+FVIII deficiency patients after surgical procedures. Evaluation of effectiveness and possible adverse events. Methods: We described 3 patients with combined mild-moderate FV+FVIII deficiency. Characteristics of the patients are described in Table 1. There were 8 relevant surgical procedures between 2014 and 2021 performed with previous administration of frozen fresh plasma and/or FVIII replacement. Antifibrinolytic agents were used depending on the patient comorbidities and surgical site. Characteristics of procedures are described in Table 2. Table 1 - Age at diagnosis (years) Gender FV FVIII Gene(s) involved PATIENT 1 2 Female 8-12% 12-17% Homozigous for LMAN1 mutation PATIENT 2 51 Male 13-15% 29-31% Homozigous for LMAN1 mutation PATIENT 3 Childhood onset Male 14-20% 25-29% Unknown Results: No relevant bleeding events were documented and all procedures were successfully completed. FFP dosage has been adapted not only to the FV level but also to patient comorbidities to avoid congestive cardiac symptoms. Desmopressin has only been used in one patient for a minor procedure. Image:Summary/Conclusion: FFP and FVIII factor replacement is a safe measure for periprocedural preparation, and the correspondent dose can be adapted to patient characteristics including bleeding phenotype, comorbidities and surgical site without increasing hemorrhagic risks. Desmopressin maybe underused.
P1541: PRELIMINARY RESULTS OF TWO YEARS FOLLOW-UP OF TYPE 1 GAUCHER DISEASE PATIENTS TREATED WITH ELIGLUSTAT IN TRAZELGA PROJECT.
Authors: I. Serrano-Gonzalo, L. López de Frutos, I. Arévalo-Vargas, M. Á. Fernández-Galán, M. Morales-Conejo, M. V. Calle-Gordo, I. Vitoria-Miñana, P. Correcher-Medina, A. Madinaveitia-Ochoa, J. Á. Hernández-Rivas, J. García-Frade, D. Ibarretxe-Gerediaga, F. Delgado-Mateos, M. D. M. Tormo-Díaz, M. Á. Ruíz-Guinaldo, E. Mora-Casterá, M. S. Noya-Pereira, A. Albarracín-Arraigosa, M. López-Dupla, J. Balanzat-Muñoz, M. L. Lozano-Almela, F. Labbadia, M. Andrade-Campos, P. Giraldo
Journal: HemaSphere (2022)

Background: Eliglustat (Cerdelga®, Sanofi-Genzyme), is a substrate reduction therapy approved in European Union in 2015 for Gaucher disease type-1 (GD1) adult patients. TRAZELGA (GEE-ELI-2017-01) project is a prospective follow-up post-authorization study designed as a tool to uniformly study the response to eliglustat therapy. Aims: In this study, it is intending to determine if the drug administration maintains or improve clinical goals and/or plasmatic biomarkers as well as to evaluate eliglustat therapy response in terms of safety, effectiveness and patient´s satisfaction. Methods: Clinical parameters, image bone assessment, and plasmatic biomarkers were analyzed for two years on follow-up: chitotriosidase [ChT] activity was evaluated by fluorimetry, glucosylsphingosine [GluSph] concentrations by LC-MSMS and CCL18/PARC, C5a, C3, YKL-40, Cathepsin S, Hepcidin and Lipocalin concentrations by immunoquantification assay. A quality of life study (SF-36), disease severity score index evaluation (GD-DS3) and compilation of side effects are included. Non-parametric statistical analysis was performed and p<0.05 was considered as statistically significant. Results: 29 Spanish GD1 patients were included, 16 were males and 13 female. The median age was 42.0 (30.00-55.50) years. All patients were treated before starting eliglustat: 26 with enzyme replacement therapy, 2 with substrate reduction therapy and 1 with a combined therapy. The median age of treatment before eliglustat was 17.0 (10.00-21.50). A statistically significant decrease on GluSph and CCL18/PARC (p=0.0393 and p=0.0019, respectively) was observed meanwhile ChT was maintain. Clinical data and the analysis of the rest of the biomarkers is going on. Summary/Conclusion: Eliglustat mantains or improve biochemical biomarkers after two years treatment. Disclosure: TRAZELGA (GZ-2017-11713) received a grant from Sanofi Genzyme for its realization.
PB1939: CLINICAL AND GENETIC CHARACTERISTICS OF PATIENTS WITH GAUCHER DISEASE: A SINGLE CENTER EXPERIENCE
Authors: S. Elsayed, R. Shawky, A. Abdelkalek
Journal: HemaSphere (2022)

Background: Gaucher disease is a rare lysosomal storage disease due to deficiency of β-glucosidase and autosomal recessive inheritance. Clinical presentation includes organomegaly, skeletal and variable neurological manifestations with two main types: neuropathic type I and non-neuropathic type 3. Aims: To describe the main clinical and genetic features of Gaucher patients (GD) in the Genetics Department, Ain Shams University Methods: The study included 27 patients with Gaucher disease confirmed by both enzyme and molecular testing. History, clinical data and laboratory and imaging investigations were brought from patients’ files. Results: Patients ages ranged between 3 months and 41 years. Male to female ratio was 1.7:1. Their age ranged between 3 months and 41 years. Parental consanguinity was present in 81.5 %. Two thirds of patients were type III. Age at presentation was significantly lower in type III patients (1.6±2.4 years) compared to 14.3±12.3 in patients with type 1. The most common presenting symptom was abdominal distention (96.3%) followed by developmental delay, while growth retardation was the least. Huge splenomegaly and low hemoglobin level was significantly present in type III, while thrombocytopenic platelet level showed no significant difference between the two types. The most common pathogenic variant was L444P present in homozygous state in all type III patients. All patients received enzyme replacement therapy (ERT) with improvement of growth, organomegaly, cytopenias and severity score and limited improvement of neurological manifestations. Mortality was present in 12 patients (due to pneumonia), all were type III. Summary/Conclusion: Although beta-thalassemia is the most common cause of chronic anemia in Egypt especially if present with organomegaly, the presence of thrombocytopenia should alert the physician to exclude GD. ERT is effective in improving visceromegaly with limited effect on neurological manifestations. Type III GD is the most common type in Egypt with one common pathogenic variant which could be used as tool of national wide carrier screening program.
Abstract LB210: Preclinical development and evaluation of allogeneic CAR T cells targeting CD70 for the treatment of renal cell carcinoma
Authors: Siler Panowski, Surabhi Srinivasan, Nguyen Tan, Silvia Tacheva-Grigorova, Bryan Smith, Yvonne Mak, Hongxiu Ning, Jonathan Villanueva, Dinali Wijewarnasuriya, Shanshan Lang, Zea Melton, Adit Ghosh, Mathilde Dusseaux, Roman Galetto, Jonathan Heyen, Tao Sai, Thomas Van Blarcom, Javier Chaparro-Riggers, Barbra Sasu
Journal: Cancer Research (2022)

Abstract CD70 is highly expressed on renal cell carcinoma (RCC), with limited normal tissue expression, making it an attractive CAR T target for an immunogenic solid tumor indication. Here we generated and characterized a panel of anti-CD70 scFv-based CAR T cells. Despite the expression of CD70 on T cells, production of CAR T from a subset of scFvs with potent in vitro activity was achieved. Expression of CD70 CARs was found to mask CD70 detection in cis and provide protection from CD70 CAR T-mediated fratricide. Two unique classes of CAR T cells were identified with differing memory phenotype, activation status, and cytotoxic activity. Epitope mapping revealed CARs binding to the membrane distal region of the CD70 extracellular domain (ECD) fall into the more active and differentiated class, as compared to CARs binding the membrane proximal region of the CD70 ECD. CD70 CAR T cells were evaluated with rituximab-based off switches to provide control over CAR T function and displayed robust antitumor activity against RCC cell lines and patient-derived xenografts in mouse models. Tissue cross reactivity studies to evaluate off-target binding with two lead CARs showed membrane staining in rare tissue-resident lymphocytes, thus matching the known expression pattern of CD70. Expected findings related to T cell activation, and elimination of CD70-expressing cells were observed in a cynomolgus monkey CD3-CD70 bispecific toxicity study and included cytokine release and loss of cellularity in lymphoid tissues. Lastly, highly functional CD70 allogeneic CAR T cells were produced at large scale through elimination of the T cell receptor by TALEN® gene editing. Taken together, these efficacy and safety data support the evaluation of CD70 CAR T cells for the treatment of RCC and led to the advancement of an allogeneic CD70 CAR T candidate into Phase I clinical trials. Citation Format: Siler Panowski, Surabhi Srinivasan, Nguyen Tan, Silvia Tacheva-Grigorova, Bryan Smith, Yvonne Mak, Hongxiu Ning, Jonathan Villanueva, Dinali Wijewarnasuriya, Shanshan Lang, Zea Melton, Adit Ghosh, Mathilde Dusseaux, Roman Galetto, Jonathan Heyen, Tao Sai, Thomas Van Blarcom, Javier Chaparro-Riggers, Barbra Sasu. Preclinical development and evaluation of allogeneic CAR T cells targeting CD70 for the treatment of renal cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr LB210.